Background: The decline in male sperm production capacity may be related not only to the testes themselves but also may be indirectly influenced by other tissues outside the testes. However, the association between extragonadal tissues and testicular function has not been systematically investigated. Methods: Using the public Genotype-Tissue Expression database (GTEx) bulk RNA-seq dataset, we stratified GTEx subjects into testis functional "good" and "poor" groups based on the expression profiles of spermatogenesis-related genes in testicular tissues. To identify the influence of extragonadal tissues on male fertility, we then compared RNA-seq data of various non-testicular tissues between these two groups. Results: Our findings reveal significant associations between testicular function and the pituitary gland, adrenal gland, lungs, liver, and key brain regions involved in neuroendocrine regulation, including the amygdala, substantia nigra, anterior cingulate cortex, and frontal cortex. Conclusions: These results suggest potential extragonadal pathways that may be associated with male fertility, and provide potential candidate biomarkers for testicular dysfunction diagnosis.

Development of inhibitory antibodies (inhibitors) against factor VIII (FVIII) is a significant complication of protein replacement therapy in hemophilia A (HA). Platelets (Plt), traditionally viewed as mediators of hemostasis, also modulate immune responses through cytokine release and interactions with immune cells. Harnessing these immunomodulatory properties may provide a novel strategy to prevent or suppress inhibitor formation. The object was to investigate whether FVIII-engineered Plt and related Plt-based products modulate FVIII immune responses in HA mice. FVIII-containing Plt were isolated from 2bF8 transgenic mice. FVIII-deficient mice were infused with intact FVIII-containing Plt, desialylated FVIII-containing Plt (dPlt), or acidified Plt lysates in combination with or before recombinant human FVIII (rhF8) exposure. Anti-FVIII antibody titers were determined by the Bethesda assay and enzyme-linked immunosorbent assay, and T-cell responses were analyzed by flow cytometry and proliferation assays. Co-infusion of FVIII Plt with rhF8 significantly reduced inhibitor titers compared with rhF8 alone. Acidified FVIII Plt lysates were potent, decreasing inhibitor titers by >20-fold when co-infused with rhF8. In contrast, co-infusion of dPlt with rhF8 did not suppress immune responses. However, repeated pre-sensitization with dPlt alone promoted immune tolerance to FVIII, evidenced by reduced inhibitor titers upon rhF8 immunization and attenuated CD4⁺ T-cell proliferation upon subsequent rhF8 exposure. These findings reveal a hierarchy of immune modulation, with intact Plt providing partial protection, lysates strongly suppressing immune responses, and dPlt inducing immune tolerance. FVIII-engineered Plt and Plt-derived products are potent immune modulators. These strategies offer novel and translatable approaches to both restore hemostasis and prevent or eradicate inhibitors in HA.

During oocyte growth, rRNAs are transcribed in nucleolus and participate in ribosome formation, but dynamic changes of rRNAs during oocyte maturation haven't yet been studied. Using RNA-FISH, we found rRNAs in germinal vesicle breakdown (GVBD) stage oocytes and a small number of oocytes at the pro-metaphase of the first meiosis stage still associate with chromosomes, and can be labeled with 5-Ethynyl Uridine (5-EU). In oocytes, rDNA transcription factor UBTF are present within nucleoli at germinal vesicle (GV) stage or near chromosomes at GVBD stage. However, RNA polymerase I can only be found at GV but not GVBD oocytes. In contrast, RNA helicase DDX3X doesn't colocalize with UBTF in GV oocytes but does colocalize with UBTF in GVBD oocytes. During oocyte maturation, the proportion of oocyte with DDX3X focus gradually decrease after GVBD. At GVBD stage, both UBTF and DDX3X are associated with rRNA condensates. When treating oocytes with DDX3X inhibitor RK-33, the decrease of 5-EU signals in oocytes will be delayed. Both treatment of RK-33 or microinjection of mutated human DDX3X cRNAs significantly reduce polar body extrusion rate and increase rate of spindle abnormalities in oocytes. Based on these findings, we hypothesize that rRNA accumulated during GV stage may entangle condensed chromosomes during GVBD and impairs chromosome segregation, and RNA helicases such as DDX3X are recruited to rRNA condensate to facilitate its clearance, thereby promoting proper chromosome segregation in oocytes. These data reveal a novel mechanism regulating chromosome segregation in oocytes.

OBJECTIVE:To investigate the molecular alterations of splenocytes associated with anti-factor Ⅷ (FⅧ) immune response and the underlying mechanisms based on hemophilia A (HA) murine model via RNA sequencing (RNA-seq) technology. METHODS:Severe HA mice were immunized with recombinant human factor Ⅷ (rhF8) weekly for 4 weeks to establish an FⅧ inhibitor model. High quality raw data were obtained by using bulk RNA-seq and CASAVA base identification technology, and the differentially expressed genes (DEGs) were identified. The DEGs were statistically classified by gene ontology (GO) annotation to obtain information on the major signaling pathways and biological processes involved in anti-FⅧ immune response in HA mouse splenocytes. The cell clusters, genes, and signaling pathway datasets were comprehensively analyzed by GO, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and single cell RNA-seq (ScRNA-seq) analysis, respectively. Flow cytometry analysis was used to verify the changes in T follicular helper cells (Tfh) and regulatory T cells (Treg). RESULTS:A total of 3731 DEGs was identified, including 2275 genes with up-regulated expression and 1456 genes with down-regulated expression. The DEGs were enriched in helper T cell differentiation, cytokine receptor, T cell receptor signaling pathway, ferroptosis, etc. Uniform Manifold Approximation and Project (UMAP) downscaling and visualization analysis yielded a total number of 11 T/NK cell subsets, visualizing the overall expression distribution of C-X-C chemokine-specific receptor gene cxcr5 among these T/NK cell subsets. Higher expression of cxcr5 was found in activated Tfh from FⅧ inhibitor mice, in comparison to the control group. The visualization using Upset plot R language showed a close interaction between Tfh and Treg. Moreover, the increased frequencies of Tfh and the decreased frequencies of Treg in inhibitor mouse splenocytes were further verified by flow cytometry analysis. CONCLUSION:Multiple immune cell subsets, signaling pathways, and characteristic genes may be involved in the process of anti-FⅧ immune response in HA mouse splenocytes. The molecules involved in the regulation of Tfh/Treg may play key roles, which provide potential biological targets and therapeutic strategies for HA patients with inhibitors in the future.

Background: The development of inhibitory antibodies against factor VIII (FVIII) remains a major complication of replacement therapy in patients with hemophilia A (HA). Our previous work demonstrated that platelet-targeted FVIII gene therapy can induce immune tolerance. However, transfusion of FVIII-containing platelets alone neither triggers inhibitor formation nor leads to consistent immune tolerance induction. Objective: This study investigates the immunomodulatory potential of FVIII-containing platelets and their derivatives in shaping immune responses to FVIII in HA mice. Methods: Human B-domain deleted FVIII was expressed in platelets of 2bF8 transgenic (2bF8Tg) mice under the platelet-specific αIIb promoter. Platelets were harvested and used intact, enzymatically desialylated (dPlts), or lysed under acidic conditions. These products were infused into FVIIInull mice either alone or with recombinant human FVIII (rhFVIII), followed by immunization. Inhibitor titers were measured using the Bethesda assay. Regulatory T (Treg) induction and CD4⁺ T-cell proliferation were also evaluated. Results: Co-infusion of 2bF8Tg platelets with rhFVIII significantly reduced FVIII inhibitor titers by 8-fold compared to rhFVIII alone (32 ± 36 vs. 253 ± 125 BU/mL). Co-administration of acidified 2bF8Tg platelet lysates, which efficiently induce functional Treg cells, further decreased inhibitor titers by 21-fold. Desialylation of GPIbα removed 98% of terminal sialic acids, producing apoptotic-like 2bF8Tg-dPlts. Infusion of these dPlts expanded Treg populations in HA mice without inducing inhibitors. Moreover, prior sensitization with 2bF8Tg-dPlts before rhFVIII immunization markedly lowered both the incidence and titers of inhibitors. Splenic CD4⁺ T cells from dPlt-sensitized, rhFVIII-immunized mice failed to proliferate upon in vitro rhFVIII restimulation, whereas control animals showed a 5-fold increase in dividing CD4⁺ T cells under the same conditions. Conclusion: FVIII-containing platelets—especially in the form of desialylated platelets or acidified lysates—effectively suppress inhibitor formation and promote immune tolerance. This novel immune modulatory strategy may represent a translational approach to prevent or treat FVIII inhibitor development in hemophilia A. Keywords: Hemophilia A, Immune tolerance, Factor VIII, Platelets, Inhibitors

Declining of male fertility may stem from intrinsic testicular dysfunction or systemic influences from other tissues. However, beyond the hypothalamic-pituitary-testis axis, little is known about which organs interact with the testes or whether they affect spermatogenic capacity. To investigate this, we analyzed GTEx open-access data, comparing transcriptomic profiles across various tissues in men with high and low testicular spermatogenic activity. Our findings reveal significant associations between testicular function and the pituitary gland, adrenal gland, lungs, liver, and key brain regions involved in neuroendocrine regulation, including the amygdala, substantia nigra, anterior cingulate cortex, and frontal cortex. These results suggest potential extragonadal pathways that modulate male fertility, highlighting candidate biomarkers for testicular dysfunction and novel therapeutic targets for infertility intervention. ### Competing Interest Statement The authors have declared no competing interest.

The development of anti-factor VIII (FVIII) neutralizing antibodies (inhibitors) remains challenging complication in hemophilia A (HA) patients undergoing prophylactic FVIII replacement therapy. The pathogenesis of FVIII inhibitor formation remains unclear. Chemokine CXCL13, a key ligand for follicular helper T cells (TFHs), in the context of inhibitor development were assessed in the present study. A total of 113 HA patients, with and without inhibitors, along with 72 healthy volunteers, were enrolled. Results demonstrated abnormally elevated levels of CXCL13 in HA patients, with a 2.0-fold increase in patients with inhibitors compared to those without. Similarly, CXCL13 levels were significantly elevated in both wild-type and HA mice with FVIII inhibitors. The proportions of circulating and splenic TFHs were markedly higher in inhibitor patients and murine models and positively correlated with CXCL13 levels. Moreover, plasma levels of B cell activating factor and the inflammatory biomarker HMGB1 were significantly increased in both human and animal inhibitor cohorts. An increased frequency of germinal center B cells was observed in splenocytes from inhibitor mice. In vitro study revealed human dermal microvascular endothelial cells undergoing immunogenic ferroptosis when conditioned with high levels of CXCL13, which was associated with down-regulation of ferroptosis suppressors SLC7A11 and GPX4, activation of the Nrf2 pathway, and increased intracellular reactive oxygen species. The findings of this study suggest that CXCL13 play a pivotal role in the microenvironment of anti-FVIII antibody development. Targeting CXCL13 may offer a potential therapeutic approach for FVIII inhibitors in HA.

Background: Platelet gene therapy is effective in hemophilia A (HA) mice even with inhibitors. Fludarabine (Flu), along with busulfan (Bu) or melphalan (Mel), preconditioning has been shown to be highly effective for hematopoietic stem cell transplantation in the clinic. Objectives: To evaluate the efficacy of Bu-Flu and Mel-Flu preconditioning in platelet gene therapy of HA with inhibitors. Methods: Bu-Flu and Mel-Flu were used to condition HA mice preimmunized with recombinant human factor (F)VIII. An optimal 660 centigray total body irradiation was used as a control regimen in parallel. Platelet-FVIII expression was introduced by transplantation of 2bF8 lentivirus (LV)-transduced hematopoietic stem cells. Animals were analyzed by fluorescence-activated cell sorting, quantitative polymerase chain reaction, FVIII assays, and tail bleeding tests. Results: Bu-Flu, but not Mel-Flu, enabled successful 2bF8 gene therapy. All recipients achieved >55% chimerism post hematopoietic stem cell transplantation in both Bu-Flu and 660 centigray groups, with comparable copy numbers of 2bF8 cassette and the platelet-FVIII levels. The bleeding phenotype was rescued in 2bF8LV-transduced recipients. FVIII inhibitor titers declined with time, with comparable disappearance time of inhibitors between the 2 groups. When animals were rechallenged with recombinant human FVIII after the titers dropped to undetectable levels, no inhibitors were detected in 2bF8LV-transduced recipients. In contrast, all untransduced transplanted control mice produced inhibitors. These data demonstrate that immune tolerance was established in 2bF8LV-transduced primed HA mice under Bu-Flu conditioning. Conclusion: Bu-Flu preconditioning allows for successfully introducing platelet-FVIII expression to restore hemostasis and induce immune tolerance in primed HA mice, suggesting that this approach is a promising clinically translatable strategy for gene therapy of HA with inhibitors.

Background Immune thrombocytopenia (ITP) is an autoimmune bleeding disorder defined by a diminished platelet count. ITP pathogenesis involves intricate changes to both cellular and humoral immunity. The pivotal roles of follicular helper T (TFH) cells in the maturations of B cells and the production of antibodies are well-established. However, the specific role of TFH to the immunopathogenesis of ITP remain incompletely understood. This study aimed to clarify the association of CXCL13/CXCR5 axis with TFH in adults with ITP. Methods A total of 97 ITP patients and 41 healthy controls were enrolled. CD4+CXCR5+ TFH, CD4+CXCR5+PD-1+ TFH, CD4+CXCR5+Foxp3+ follicular regulatory T cells (TFR), and desialylated platelets in peripheral blood were measured by flow cytometry. Plasma cytokines were assessed by enzyme-linked immunosorbent assay. CD4+ T cells cocultured with chemokine CXCL13 in vitro was performed for the measurement of TFH proliferation. Intracellular production of reactive oxygen species (ROS) was examined by dichlorodihydrofluorescein diacetate (DCFH-DA) probe staining. Results We observed a significant increase in circulating TFH and a marked decrease in circulating TFR in the entire ITP cohort. The ratio of TFH/TFR was elevated, accompanied by heightened levels of platelet desialylation, cytokines BAFF, HMGB1, and IL-21, while levels of IL-10 were downregulated in adults with ITP. Notably, patients with ITP exhibiting platelet count below 50 × 109/L had dramatically elevated levels in both chemokine CXCL13 and its receptor CXCR5+ TFH compared to those with platelet count above 100 × 109/L. High frequencies of TFH correlated with poor therapeutic response. Furthermore, in vitro CD4+ T cell proliferation assay demonstrated a CXCL13 dose-dependent increase in the frequencies in both CD4+CXCR5+ TFH and CD4+CXCR5+PD-1+ TFH from ITP patients. Intriguingly, DCFH-DA assay illustrated a significant enhancement in intracellular ROS generation in CXCR5+ T cell subsets, especially in CD4+CXCR5+PD-1+ TFH from 4 patients with ITP. Conclusions These results underscore the pivotal role of CXCL13/CXCR5 axis-drived TFH expansion in the pathogenesis of ITP, providing a potential disease severity biomarker.

Anti-factor VIII (FVIII) antibody development poses a significant challenge in hemophilia A (HA) patients receiving FVIII protein replacement therapy. There is an urgent need for novel therapeutic strategies to inhibit the production of anti-FVIII inhibitory antibodies (inhibitors) in HA. This study aimed to investigate a combination monoclonal antibody (mAb) therapy targeting CXCL13 and CD20 on the development of anti-FVIII antibodies in a HA murine model, along with the underlying mechanisms involved. Specifically, mAbs targeting mouse CD20 (18B12) with an IgG2a backbone and mouse CXCL13 (2C4) with an IgG1 backbone were synthesized. HA mice with FVIII inhibitors were established, and the results revealed that the combination therapy of anti-mCD20 with α-mCXCL13 significantly suppressed anti-FVIII antibody development and induced FVIII tolerance. Furthermore, this combination therapy led to a marked reduction of peripheral and splenic follicular helper T cells and an enhancement of regulatory T cell induction, along with sustained depletion of bone marrow and splenic plasma cells in HA mice with preexisting FVIII immunity. Thus, the concurrence of blockage of CD20 and neutralization of CXCL13 hold promise as a therapeutic strategy for HA patients with inhibitors.

Bortezomib (BTZ), a proteasome inhibitor, is a promising therapeutic option for multiple myeloma (MM) patients. However, drug resistance often occurs, leading to disease relapse and poor prognosis. In this study, we aimed to identify novel genes associated with drug resistance and investigate their roles in BTZ resistance. Through the screening of 26 genes frequently associated with chemosensitivity or drug resistance, we discovered that KDM4C, a histone demethylase, exhibited increased expression in BTZ-resistant MM cells compared to their sensitive counterparts. Overexpression of KDM4C enhanced the tolerance of a MM cell line to the drug, whereas the knockdown of KDM4C, using shRNA, increased the sensitivity of resistant cells to BTZ treatment. This suggests that KDM4C plays a pivotal role in conferring BTZ resistance. Our study offers fresh insights into BTZ resistance in MM and highlights KDM4C as a potential target for overcoming drug resistance.

Acute myeloid leukemia (AML) is an aggressive hematopoietic malignancy with poor prognosis and high recurrence rate. The discovery of more effective therapeutic strategies for AML plays a crucial role. The present work showed that E35, a novel derivative of emodin, significantly inhibited cell proliferation and induced autophagy and apoptosis in AML cells. Treatment with E35 markedly induced Beclin-1, LC3-II, cleaved Caspase-9 and PARP, and suppressed mitogen-activated protein kinase (MAPK) pathway. E35 exposure evoked autophagic activity prior to apoptosis induction, and autophagy inhibition by 3-methyladenine (3-MA) dramatically increased E35-induced apoptosis in both AML cell lines and patient-derived AML cells. Nevertheless, study on AML xenograft model showed that the combination E35 with 3-MA exhibited much more inhibitory effects on leukemia cell growth in vivo. No obvious adverse reactions occurred in the xenograft animals administered E35 alone or its cotreatment with 3-MA. These findings suggest that E35 could exert anti-leukemia effects, and that the combination of E35 and autophagy inhibitor might prove a more highly efficient strategy for AML treatment.

Platelets are small, versatile blood cells that are critical for hemostasis/thrombosis. Local platelet accumulation is a known contributor to proinflammation in various disease states. However, the anti-inflammatory/immunosuppressive potential of platelets has been poorly explored. Here, we uncovered, unexpectedly, desialylated platelets (dPLTs) down-regulated immune responses against both platelet-associated and -independent antigen challenges. Utilizing multispectral photoacoustic tomography, we tracked dPLT trafficking to gut vasculature and an exclusive Kupffer cell-mediated dPLT clearance in the liver, a process that we identified to be synergistically dependent on platelet glycoprotein Ibα and hepatic Ashwell–Morell receptor. Mechanistically, Kupffer cell clearance of dPLT potentiated a systemic immunosuppressive state with increased anti-inflammatory cytokines and circulating CD4+ regulatory T cells, abolishable by Kupffer cell depletion. Last, in a clinically relevant model of hemophilia A, presensitization with dPLT attenuated anti-factor VIII antibody production after factor VIII ( infusion. As platelet desialylation commonly occurs in daily-aged and activated platelets, these findings open new avenues toward understanding immune homeostasis and potentiate the therapeutic potential of dPLT and engineered dPLT transfusions in controlling autoimmune and alloimmune diseases.

Thrombocytopenia is a multifactorial condition that frequently involves concomitant defects in platelet production and clearance. The physiopathology of low platelet count in thrombocytopenia remains unclear. Sialylation on platelet membrane glycoprotein and follicular helper T cells (TFHs) are thought to be the novel platelet clearance pathways. The aim of this study was to clarify the roles of platelet desialylation and circulating TFHs in patients with immune thrombocytopenia (ITP) and non-ITP thrombocytopenia. We enrolled 190 patients with ITP and 94 patients with non-ITP related thrombocytopenia including case of aplastic anemia (AA) and myelodysplastic syndromes (MDS). One hundred and ten healthy volunteers were included as controls. We found significantly increased desialylated platelets in patients with ITP or thrombocytopenia in the context of AA and MDS. Platelet desialylation was negatively correlated with platelet count. Meanwhile, the circulating TFH levels in patients with thrombocytopenia were significantly higher than those of normal controls, and were positively correlated with desialylated platelet levels. Moreover, TFHs-related chemokine CXCL13 and apoptotic platelet levels were abnormally high in ITP patients. The upregulation of pro-apoptotic proteins and the activation of the MAPK/mTOR pathway were observed in the same cohort. These findings suggested that platelet desialylation and circulating TFHs may become the potential biomarkers for evaluating the disease process associated with thrombocytopenia in patients with ITP and non-ITP.

The development of coagulation factor VIII (FVIII) inhibitory antibodies is a serious complication in hemophilia A (HA) patients after FVIII replacement therapy. Inhibitors render regular prophylaxis ineffective and increase the risk of morbidity and mortality. Immune tolerance induction (ITI) regimens have become the only clinically proven therapy for eradicating these inhibitors. However, this is a lengthy and costly strategy. For HA patients with high titer inhibitors, bypassing or new hemostatic agents must be used in clinical prophylaxis due to the ineffective ITI regimens. Since multiple genetic and environmental factors are involved in the pathogenesis of inhibitor generation, understanding the mechanisms by which inhibitors develop could help identify critical targets that can be exploited to prevent or eradicate inhibitors. In this review, we provide a comprehensive overview of the recent advances related to mechanistic insights into anti-FVIII antibody development and discuss novel therapeutic approaches for HA patients with inhibitors.

Elevated extrachromosomal circular DNA (eccDNA) has been reported to accelerate tumor pathogenesis. Although the eccDNA profiles of other tumors have been established, the landscape of the eccDNA of acute myeloid leukemia (AML) has not been revealed. Our study first depicted the eccDNA profile of normal hematopoiesis and AML evolution by exploiting the ATAC-seq and RNA-seq data from nine healthy donors and 12 AML patients, which contained a total of 137 cell samples and 96 RNA-seq samples (including 16 blood cell types of the normal hematopoietic and AML hierarchies). We found the number of eccDNAs generally increased with the evolution of normal hematopoiesis and AML. The ecDNAs and ring chromosomes were found to reappear both in normal hematopoiesis and AML cells. Furthermore, we compared the eccDNAs of AML with normal cells. There were almost 300 AML-specific genes, including the known oncogenes NRAS, MCL1, EVI1, GATA2, WT1, and PAK1. And the ecDNA (chr11: 58668376-58826008) occurred in five out of 17 AML evolution-related cells, which was associated with the high expression of the GLYATL1 gene and the high expressed GLYATL1 was a poor prognostic factor. In conclusion, the eccDNA profiles of normal hematopoiesis and AML evolution were depicted and the recurrent eccDNAs we revealed might be utilized in the treatment of AML as biomarkers.

Immune thrombocytopenia (ITP) is an autoimmune disease characterized by immune-mediated destruction of one's own platelets. The progression of thrombocytopenia involves an imbalance of platelet production and clearance. B cells can induce autoantibodies, and T cells contribute to the pathological progression as well. Some patients with ITP have a poor response to common first-line therapies. Recent studies have shown that a novel Fc-independent platelet clearance pathway is associated with poor prognosis in these patients. By this pathway, desialylated platelets can be cleared by Ashwell-Morell receptor (AMR) on hepatocytes. Research has demonstrated that patients with refractory ITP usually have a high level of desialylation, indicating the important role of sialylation on platelet membrane glycoprotein (GP) in patients with primary immune thrombocytopenia, and neuraminidase 1(NEU1) translocation might be involved in this process. Patients with ITP who are positive for anti-GPIbα antibodies have a poor prognosis, which indicates that anti-GPIbα antibodies are associated with this Fc-independent platelet clearance pathway. Experiments have proven that these antibodies could lead to the desialylation of GPs on platelets. The T follicular helper (TFH) cell level is related to the expression of the anti-GPIbα antibody, which indicates its role in the progression of desialylation. This review will discuss platelet clearance and production, especially the role of the anti-GPIbα antibody and desialylation in the pathophysiology of ITP and therapy for this disease.

本文分析了主要慕课平台和移动学习工具特点,探索高等医学不同课程开展全线上教学的模式方法、线上教学质量等效保障措施和思政元素融入方式.思考全线上教学存在的问题,总结特殊环境下,应用"互联网+"信息化技术实施全课程线上教学改革的经验.

Acute lymphoblastic leukemia (ALL) is an aggressive malignancy. Adults with ALL have more than 50% relapse rates. We have previously validated that overexpression of nucleophosmin (NPM) is involved in the multidrug resistance (MDR) development during ALL; and a synthetically engineered recombinant NPM binding protein (NPMBP) has been developed in our group; NPMBP and doxorubicin (DOX) can be conjugated in a nanoparticle-based drug delivery system named DOX-PMs-NPMBP to counteract MDR during ALL. Here, we evaluated the antileukemia potential of DOX-PMs-NPMBP in resistant ALL cells. This study demonstrates that DOX-PMs-NPMBP significantly enhances chemosensitivity to DOX in ALL cells. Despite at variable concentrations, both resistant and primary ALL cells from relapsed patients were sensitive to DOX-PMs-NPMBP. In detail, the half maximal inhibitory concentration (IC50) values of DOX-PMs-NPMBP were between 1.6- and 7.0-fold lower than those of DOX in cell lines and primary ALL cells, respectively; and apoptotic cells ratio was over 2-fold higher in DOX-PMs-NPMBP than DOX. Mechanistically, p53-driven apoptosis induction and cell cycle arrest played essential role in DOX-PMs-NPMBP-induced anti-leukemia effects. Moreover, DOX-PMs-NPMBP significantly inhibited tumor growth and prolonged mouse survival of ALL xenograft models; and no systemic toxicity occurrence was observed after treatment during follow-up. In conclusion, these data indicate that DOX-PMs-NPMBP may significantly exert growth inhibition and apoptosis induction, and markedly improve DOX antileukemia activity in resistant ALL cells. This novel drug delivery system may be valuable to develop as a new therapeutic strategy against multidrug resistant ALL.